Age-related Changes in Trigeminal Ganglion Macrophages Enhance Orofacial Ectopic Pain After Inferior Alveolar Nerve Injury.

BACKGROUND/AIM: The ectopic pain associated with inferior alveolar nerve (IAN) injury has been reported to involve macrophage expression in the trigeminal ganglion (TG). However, the effect of age-related changes on this abnormal pain conditions are still unknown. This study sought to clarify the involvement of age-related changes in macrophage expression and phenotypic conversion in the TG and how these changes enhance ectopic mechanical allodynia after IAN transection (IANX). MATERIALS AND METHODS: We used senescence-accelerated mouse (SAM)-prone 8 (SAMP8) and SAM-resistance 1 (SAMR1) mice, which are commonly used to study ageing-related changes. Mechanical stimulation was applied to the whisker pad skin under light anaesthesia; the mechanical head withdrawal threshold (MHWT) was measured for 21 d post-IANX. We subsequently counted the numbers of Iba1 (macrophage marker)-immunoreactive (IR) cells, Iba1/CD11c (M1-like inflammatory macrophage marker)-co-IR cells, and Iba1/CD206 (M2-like anti-inflammatory macrophage marker)-co-IR cells in the TG innervating the whisker pad skin. After continuous intra-TG administration of liposomal clodronate Clophosome®-A (LCCA) to IANX-treated SAMP8-mice, the MHWT values of the whisker pad skin were examined. RESULTS: Five days post-IANX, the MHWT had significantly decreased in SAMP8 mice compared to SAMR1-mice. Iba1-IR and Iba1/CD11c-co-IR cell counts were significantly increased in SAMP8 mice compared to SAMR1 mice 5 d post-IANX. LCCA administration significantly restored MHWT compared to control-LCCA administration. CONCLUSION: Ectopic mechanical allodynia of whisker pad skin after IANX is exacerbated by ageing, which involves increases in M1-like inflammatory macrophages in the TG.

Age-Related Differences in Transient Receptor Potential Vanilloid 1 and 2 Expression Patterns in the Trigeminal Ganglion Neurons Contribute to Changes in the Palatal Mucosal Heat Pain Sensitivity.

Aging affects various sensory functions of the body. However, the effect on the oral mucosal nociception has remain unclear, so this elucidation is very important. Therefore, this study aimed to evaluate the effect of age-related changes in transient receptor potential vanilloid 1 (TRPV1) and TRPV2 expression in the trigeminal ganglion (TG) neurons on intraoral mucosal heat sensitivity in the senescence-accelerated mouse prone 8 (SAMP8) model. We used 23-week-old (aged) and 7-week-old (young) SAMP8 mice. Heat stimulation was applied to the palatal mucosa under light anesthesia; moreover, the heat head withdrawal threshold (HHWT) was measured. We counted the number of TRPV1-immunoreactive (IR) and TRPV2-IR TG neurons innervating the palatal mucosa. Additionally, we investigated changes in HHWT when TRPV1 or TRPV2 antagonists (SB366791 or Tranilast) were administered to the palatal mucosa. Aged SAMP8 mice showed a higher HHWT than young SAMP8 mice. Compared with the aged SAMP8 mice, young SAMP8 mice showed a larger number of TRPV1-IR small-diameter neurons and a smaller number of TRPV2-IR medium-sized neurons innervating the palatal mucosa. SB366791 administration increased the HHWT in young, but not aged SAMP8 mice. Contrastingly, Tranilast administration increased the HHWT in aged, but not young SAMP8 mice. These results suggest that the modulation of heat pain sensitivity in the oral mucosa due to aging is dependent on changes in the TRPV1 and TRPV2 expression patterns in the TG neurons innervating the palatal mucosa.

Oxytocin-Dependent Regulation of TRPs Expression in Trigeminal Ganglion Neurons Attenuates Orofacial Neuropathic Pain Following Infraorbital Nerve Injury in Rats.

We evaluated the mechanisms underlying the oxytocin (OXT)-induced analgesic effect on orofacial neuropathic pain following infraorbital nerve injury (IONI). IONI was established through tight ligation of one-third of the infraorbital nerve thickness. Subsequently, the head withdrawal threshold for mechanical stimulation (MHWT) of the whisker pad skin was measured using a von Frey filament. Trigeminal ganglion (TG) neurons innervating the whisker pad skin were identified using a retrograde labeling technique. OXT receptor-immunoreactive (IR), transient receptor potential vanilloid 1 (TRPV1)-IR, and TRPV4-IR TG neurons innervating the whisker pad skin were examined on post-IONI day 5. The MHWT remarkably decreased from post-IONI day 1 onward. OXT application to the nerve-injured site attenuated the decrease in MHWT from day 5 onward. TRPV1 or TRPV4 antagonism significantly suppressed the decrement of MHWT following IONI. OXT receptors were expressed in the uninjured and Fluoro-Gold (FG)-labeled TG neurons. Furthermore, there was an increase in the number of FG-labeled TRPV1-IR and TRPV4-IR TG neurons, which was inhibited by administering OXT. This inhibition was suppressed by co-administration with an OXT receptor antagonist. These findings suggest that OXT application inhibits the increase in TRPV1-IR and TRPV4-IR TG neurons innervating the whisker pad skin, which attenuates post-IONI orofacial mechanical allodynia.

Aging-Related Phenotypic Conversion of Medullary Microglia Enhances Intraoral Incisional Pain Sensitivity.

Activated microglia involved in the development of orofacial pain hypersensitivity have two major polarization states. The aim of this study was to assess the involvement of the aging-related phenotypic conversion of medullary microglia in the enhancement of intraoral pain sensitivity using senescence-accelerated mice (SAM)-prone/8 (SAMP8) and SAM-resistant/1 (SAMR1) mice. Mechanical head-withdrawal threshold (MHWT) was measured for 21 days post palatal mucosal incision. The number of CD11c-immunoreactive (IR) cells [affective microglia (M1)] and CD163-IR cells [protective microglia (M2)], and tumor-necrosis-factor-α (TNF-α)-IR M1 and interleukin (IL)-10-IR M2 were analyzed via immunohistochemistry on days 3 and 11 following incision. The decrease in MHWT observed following incision was enhanced in SAMP8 mice. M1 levels and the number of TNF-α-IR M1 were increased on day 3 in SAMP8 mice compared with those in SAMR1 mice. On day 11, M1 and M2 activation was observed in both groups, whereas IL-10-IR M2 levels were attenuated in SAMP8 mice, and the number of TNF-α-IR M1 cells increased, compared to those in SAMR1 mice. These results suggest that the mechanical allodynia observed following intraoral injury is potentiated and sustained in SAMP8 mice due to enhancement of TNF-α signaling, M1 activation, and an attenuation of M2 activation accompanying IL-10 release.

Influence of power density on polymerization behavior and bond strengths of dual-cured resin direct core foundation systems.

This study examined the influence of power density on dentin bond strength and polymerization behavior of dual-cured direct core foundation resin systems. Two commercially available dual-cured direct core foundation resin systems, Clearfil DC Core Automix with Clearfil DC Bond and UniFil Core with Self-Etching Bond, were studied. Bovine mandibular incisors were mounted in autopolymerizing resin and the facial dentin surfaces were ground wet on 600-grit SiC paper. Dentin surfaces were treated according to manufacturer's recommendations. The resin pastes were condensed into the mold and cured with the power densities of 0 (no irradiation), 100, 200, 400 and 600 mW/cm2. Ten specimens per group were stored in 37 degrees C water for 24 hours, then shear tested at a crosshead speed of 1.0 mm/minute in a universal testing machine. An ultrasonic measurement device was used to measure the ultrasonic velocities through the core foundation resins. The power densities selected were 0 (no irradiation), 200, and 600 mW/cm2, and ultrasonic velocity was calculated. ANOVA and Tukey HSD tests were performed at a level of 0.05. The highest bond strengths were obtained when the resin pastes were cured with the highest power density for both core foundation systems (16.8 +/- 1.9 MPa for Clearfil DC Core Automix, 15.6 +/- 2.9 MPa for UniFil Core). When polymerized with the power densities under 200 mW/cm2, significantly lower bond strengths were observed compared to those obtained with the power density of 600 mW/cm2. As the core foundation resins hardened, the sonic velocities increased and this tendency differed among the power density of the curing unit. When the sonic velocities at three minutes after the start of measurements were compared, there were no significant differences among different irradiation modes for UniFil Core, while a significant decrease in sonic velocity was obtained when the resin paste was chemically polymerized compared with dual-polymerization for Clearfil DC Core Automix. The data suggests that the dentin bond strengths and polymerization behavior of the dual-cured, direct core foundation systems are still affected by the power density of the curing unit. With a careful choice of the core foundation systems and power density of the curing unit, the benefit of using resin composites to endodontically-treated teeth might be acceptable.

Influence of light intensity on contraction stress of flowable resins.

The purpose of this study was to evaluate the influence of power density on contraction stress of resin composite restorative materials during photo-polymerization. Six flowable resin composites, and a hybrid resin composite for comparison, were used. The composites were polymerized with the power density adjusted to either 100 or 600 mW/cm(2). Stress development was determined with a custom-made tensilometer. The adhesive was placed in a thin layer on a steel rod and resin paste was packed into the mold. The contraction force (N) generated during polymerization was continuously recorded and the maximum contraction stress (MPa) was calculated. Data were analyzed statistically. When the power density was adjusted to 100 mW/cm(2), the average contraction stress ranged from 0.30 to 0.50 MPa for the flowable composites, compared with 0.35 MPa for the hybrid composite. When the power density was adjusted to 600 mW/cm(2), the average contraction stress ranged from 0.34 to 1.00 MPa for the flowable composites and 0.69 MPa for the hybrid composite comparison. For all materials tested except Estelite Flow Quick, contraction stress increased with higher power density. The present results indicate that contraction stress during polymerization is influenced by power density and resin composite type.